LOCUS       sequence-297214-        5166 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF7a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5166)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5166)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046194
FEATURES             Location/Qualifiers
     source          1..5166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             841..858
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             859..876
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(921..939)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1290..2093
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      2176..3158
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3236..3265
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3287..3874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3886..3913)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3937..4128)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4129..4920,4921..4989))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4752..4771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4990..5092)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5093..5124)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 aaaattatcc tattcttagc attgataacc ctggctacgt gtgagcttta tcactaccag
      541 gaatgcgtta gggggacaac tgtcttgttg aaggaacctt gttcatctgg aacttacgaa
      601 ggcaattcgc catttcatcc attggctgat aacaagttcg ccttaacctg ttttagtact
      661 caattcgctt tcgcttgtcc cgacggtgtg aaacacgttt atcaattgag agcgagatcc
      721 gtaagcccaa agttatttat ccgtcaagaa gaagttcaag agttgtactc cccaattttt
      781 ctaattgtcg ccgctatcgt tttcattacc ttgtgtttca cactgaagcg aaagactgaa
      841 ctcgtgccga gaggttctca ccatcatcac catcattaaa caggcccctt ttcctttgtc
      901 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
      961 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     1021 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     1081 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     1141 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     1201 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     1261 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     1321 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     1381 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     1441 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     1501 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     1561 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     1621 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     1681 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     1741 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     1801 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     1861 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     1921 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     1981 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     2041 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     2101 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     2161 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     2221 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2281 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     2341 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     2401 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     2461 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     2521 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     2581 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     2641 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     2701 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     2761 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     2821 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     2881 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     2941 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     3001 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     3061 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     3121 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     3181 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     3241 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     3301 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3361 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     3421 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3481 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3541 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3601 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3661 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3721 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     3781 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     3841 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     3901 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     3961 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4021 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4081 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     4141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4201 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     4261 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4321 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     4381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4561 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4621 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     4681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4801 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4981 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     5041 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     5101 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     5161 gttaga
//
